活性饥饿素ELISA试剂盒
Active Ghrelin ELISA试剂盒适用多物种，可以检测人, 大鼠, 小鼠的血浆样品
检测范围：2.9 ~ 184 fmol/mL
样品数/Kit：使用同一个试剂盒，可重复测定40份样品。
样品量: 50 μL

该Active Ghrelin ELISA试剂盒基于双位点夹心酶联免疫吸附试验（ELISA）的原理测量Ghrelin的活性形式。该试剂盒不仅可以检测Octanoylated的人Ghrelin，还可以检测Octanoylated的大鼠/小鼠Ghrelin(1-28)。

Ghrelin是一种新型的生长激素释放肽，是一种刺激垂体释放生长激素的酰化肽。Ghrelin从大鼠胃中分离出来，其结构由Kenji kangawa博士（日本国家心血管中心）确定为是由28个氨基酸组成的肽。Ghrelin的Ser3残基被正辛酸（n-octanoic acid）修饰，这是激素活性所必需的修饰。

试剂盒组分：

检测程序：

Pre-warm all reagents to room temperature prior to setting up the assay.

Do not dry up the wells during a measurement.

1. 150 μL of assay buffer is poured into the testing well. 50 μL of samples and standards are added into the each well. As a "Blank", 50 μL of assay buffer is added into the testing well. Then shake the plate gently. Manufacturer recommends to test duplicate for each sample. Plate is covered with transparent sheet and is incubated for 2 hours at RT.

2. Aspirate samples from wells and wash by washing buffer for 3 times. Washing buffer volume:

350 μL. Keep 1 min of interval before removing the washing buffer from wells. For removing the remnant completely, testing plate is tapped on a paper towel upside down. 200 μL of diluted HRP labeled antibody solution is poured into the wells. Testing plate is covered with transparent sheet and is incubated for 1 hour at RT.

3. Aspirate samples from wells and wash by washing buffer for 4 times. Washing buffer volume:

350 μL. Keep 1 min of interval before aspirating the washing buffer from wells. For removing the remnant completely, testing plate is tapped on a paper towel upside down. 200 μL of substrate solution is poured into each well and is incubated for 30 min at RT with shading. After the incubation, 50 μL of stopping solution is added to each well to stop reaction. Then shake the plate gently.

4. Measure the absorbance of each well at 450 nm immediately.

5. The results of unknown samples can be calculated with any computer program having a 4 (or 5)-parameter logistic function. Otherwise plot the standard concentration (X-axis) and its corresponding absorbance (Y-axis). The concentration of active ghrelin in unknown sample is determined by plotting the sample’s absorbance on the standard curve.

When the HCl is added to the samples, multiply the results by 1.1 to offset the dilution.